Structural studies on dynein-microtubule complex

动力蛋白-微管复合物的结构研究

• 批准号: 7033453
• 负责人: Masahide Kikkawa
• 金额: $ 23.55万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7033453
• 关键词: Chlamydomonas; bioenergetics; bioimaging /biomedical imaging; biophysics; calcium; cilium /flagellum motility; conformation; cryoelectron microscopy; dynein ATPase; gene mutation; high throughput technology; microtubules; nucleotides; plant genetics; protein protein interaction; protein structure function; protozoal genetics; structural biology; three dimensional imaging /topography

DESCRIPTION (provided by applicant): Microtubule-based motors, dyneins, are nano-meter scale protein machineries and convert chemical energy derived from ATP hydrolysis to mechanical movement. The mechanical movement is employed for various essential intracellular motilities, such as axonal transport, chromosome segregation, and flagellar motility. The importance of dynein functions is underlined by several dynein-related diseases, such as Kartagener's syndrome and amyotrophic lateral sclerosis (ALS). However, little is known about how dynein mutations cause the primary cilia diskinesia or even how dyneins convert chemical energy to mechanical movement. Our long-term goal is to understand the structural basis of energy conversion by dynein and its regulation. To this end we will characterize the structure of dynein-microtubule complexes using cryo-electron microscopy and three-dimensional reconstruction. We have recently obtained two initial structures of dynein-microtubule complexes. One is a low resolution structure of the outer arm dynein-microtubule complex, and another is a medium resolution structure of the microtubule binding domain of dynein, the dynein stalk. Our immediate goals are to refine the structure of the outer arm dynein-microtubule complex to 25Angstroms resolution, and to extend the structure of the dynein stalk structure to 10Angstroms resolution. By studying several structures of these complexes in functionally important states, we would like to elucidate conformational changes essential to the energy conversion, motility and regulation of dynein.

描述（由申请人提供）：基于微管的电动机，动力蛋白，是纳米尺度尺度蛋白机械，并将ATP水解衍生的化学能转化为机械运动。机械运动用于各种必需的细胞内运动，例如轴突运输，染色体分离和鞭毛运动。动力蛋白功能的重要性由几种与动力蛋白有关的疾病（例如Kartagener's综合征和肌萎缩性侧索硬化症（ALS））强调。然而，关于动力蛋白突变如何引起原发性纤毛的疾病，甚至动力蛋白如何将化学能转化为机械运动的知识知之甚少。 我们的长期目标是了解动力蛋白及其调节的能量转化的结构基础。为此，我们将使用冷冻电子显微镜和三维重建来表征动力蛋白 - 微管复合物的结构。我们最近获得了Dynein-Microubule复合物的两个初始结构。一种是外臂动力蛋白 - 微管复合物的低分辨率结构，另一个是动力蛋白茎的微管结合结构域的中等分辨率结构。我们的直接目标是将外臂动力蛋白 - 微管复合物的结构改进到25 Angstroms分辨率，并将动力蛋白茎结构的结构扩展到10angstroms分辨率。通过研究这些复合物在功能上重要的状态中的几个结构，我们想阐明构象变化对于能量转化，运动性和动力蛋白的调节至关重要。